Molecular and biological analysis of the pathogenicity of herpes simplex virus

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We analyzed the genomic DNAs of 7 substrains of the Miyama strain of HSV-1 with special reference to their neurovirulence. Five of these substrains are CPE variants (-GCr, -GCf , +GC(LPV), +GC(SPV) and +GC(81)), and two are descendent strains of -GCr (alphaV and betaV). alphaV and betaV were both recovered from persistently infected L cell cultures originally established with -GCr. DNAs of the 7 substrains were digested with BamHI and analyzed by agarose gel electrophoresis and Southern blot hybridization. Various heterogeneous fragments were detected among the 7 substrains. These fragments, however, did not seem to include or to be closely associated with genetic loci of neurovirulence. BamHI-Z fragments of all CPE variants were 1.84-1.99 Kbp, while those of both alphaV and betaV were 1.78 Kbps.DNAs extracted from virions of virulent (F) and attenuated (-GCr Miyama) strains of HSV-1 were analyzed comparatively by field-inversion gel electrophoresis. Virions of -GCr were shown to contain a DNA of about 150 Kbp, corresponding to the genomic DNA of HSV-1, and another DNA larger than the former. On the other hand, virions of F contained only the 150 Kbp DNA. However, the significance of this difference is unknown.Experimental conditions suitable for the study of latent infection of HSV-1 in mice were examined, and the following results were obtained. Double infection of mice with avirulent and virulent strains was found to be useful. Activation of latent virus in vitro was shown to be enhanced by treatment with n-butyrate. A correlation was discovered between the extent of virus growth in a neuroblastoma cell line of a particular HSV strain and the efficiency of latent infection of this virus in the trigeminal ganglion of mice.